Application of high-performance countercurrent chromatography for the isolation of steroidal saponins from Liriope plathyphylla.

High-performance countercurrent chromatography (HPCCC) with electrospray light-scattering detection was applied for the first time to isolate a spirostanol and a novel furostanol saponin from Liriope platyphylla. Due to the large differences in KD values between the two compounds, a two-step HPCCC method was applied in this study. The primary HPCCC employed methylene chloride/methanol/isopropanol/water (9:6:1:4 v/v, 4 mL/min, normal-phase mode) conditions to yield a spirostanol saponin (1). After the primary HPCCC run, the solute retained in the stationary phase (SP extract) in HPCCC column was recovered and subjected to the second HPCCC on the n-hexane/n-butanol/water system (1:9:10 v/v, 5 mL/min, reversed-phase mode) to yield a novel furostanol saponin (2). The isolated spirostanol saponin was determined to be 25(S)-ruscogenin 1-O-β-D-glucopyranosyl (1→2)-[β-D-xylopyranosyl (1→3)]-β-D-fucopyranoside (spicatoside A), and the novel furostanol saponin was elucidated to be 26-O-β-D-glucopyranosyl-25(S)-furost-5(6)-ene-1β-3β-22α-26-tetraol-1-O-β-D-glucopyranosyl (1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-fucopyranoside (spicatoside D).